Note Antidepressant-Like Effects of an Aqueous Extract of Lavender
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Food Sci. Technol. Res., 18 (3), 473–479, 2012 Note Antidepressant-like Effects of an Aqueous Extract of Lavender (Lavandula angustifolia Mill.) in Rats 1, 3 2 1 2 1 1, 4* Aya kageYaMa , Toshio ueno , Mutsumi oshio , Hideki Masuda , Hiroko hoRiuchi and Hidehiko Yokogoshi 1 Laboratory of Nutritional Biochemistry and Global COE Program, Graduate School of Nutritional and Environmental Sciences, Univer- sity of Shizuoka, Shizuoka 422-8526, Japan 2 Laboratory of Health Materials, Functional Ingredient Department, Ogawa & Co., Ltd., 15-7 Chidori Urayasushi, Chiba 279-0032, Japan 3 Present address: Division of Medical Nutrition, Tokyo Healthcare University, Tokyo 154-8568, Japan 4 Present address: Department of Food & Nutritional Sciences, Chubu University, Aichi 487-8501, Japan Received June 30, 2011; Accepted January 5, 2012 The purpose of the present study was to examine the antidepressant-like effects of an aqueous extract of lavender (LAE) using the forced swimming test (FST), the most widely used animal model of depres- sion. LAE was orally administered to rats three times, i.e., 24, 5, and 1 h prior to the FST. The administra- tion of LAE (3428 mg/kg body weight) showed a significant reduction of the immobility time in the FST, the effect of which was comparable to that of the synthetic antidepressant, imipramine (30 mg/kg). In addition, the same dose of LAE did not change the locomotor activity in the open field test. These results suggest that LAE might have antidepressant-like effects that are independent of motor stimulation. Fur- thermore, the active ingredients of LAE were suggested to be non-volatile constituents, because linalool, the main aroma constituent of lavender, was completely removed during the preparation of LAE. Possible contribution of rosmarinic acid and that of apigenin glycosides to the antidepressant-like effects of LAE were discussed on the basis of their content in LAE. Keywords: lavender, depression, antidepressant-like, forced swimming test Introduction Lavender (Lavandula angustifolia Mill.) is a famous herb Depression has become a common psychological ill- that has a long history in folk medicine and is still therapeu- ness in recent years. According to an investigation by the tically used today. The essential oil obtained by steam distil- World Health Organization International Consortium of lation from the fresh flowering tops of this plant is often used Psychiatric Epidemiology (WHO-ICPE), 6.3 − 15.7% of in aromatherapy as a relaxant (Lis-Balchin, 1996). Inhalation the world’s population has been estimated to get depression of the vapor of the lavender essential oil and its main con- once in their life (Andrade et al., 2003). Exposure to chronic stituent, linalool, has shown sedative effects in both human stress is known to be associated with a higher incidence of and animal studies (Buchbauer et al., 1991; Buchbauer et depression, though its mechanism has not been elucidated. al., 1993; Diego et al., 1998; Kuroda et al., 2005). Other Although a wide variety of antidepressant drugs are avail- pharmacological effects of this oil, including anticonvulsive able to treat depression, most of the synthetic drugs are not (Yamada et al., 1994), anxiolytic (Bradley et al., 2007), anti- without side effects. Therefore, the search for regularly eaten depressive (Seol et al., 2010), and anticonflict effects (Umezu foods with an antidepressant activity seems to be an essential et al., 2006), have also been reported. approach to finding an effective antidepressant treatment On the other hand, lavender is also used as a tea infusion without side effects. (i.e., aqueous extracts) to treat restlessness, insomnia, and nervous disorders of the stomach and intestines (Blumenthal *To whom correspondence should be addressed. et al., 1998). Furthermore, lavender contains aqueous phe- E-mail: [email protected] nolic constituents, such as hydroxycinnamic acids and fla- 474 a. kageYaMa et al. vone glycosides (Harborne and Williams, 2002), which have solt et al., 1977, Kageyama et al., 2010). Briefly, 35 male been associated with the antioxidant activities of Lamiaceae rats were divided into five groups (seven rats per group): one plants including lavender (Zheng and Wang, 2001; Dorman group served as the control, another group received imip- et al., 2004; Shan et al., 2005; Torras-Claveria et al., 2007). ramine (30 mg/kg body weight), and the other three groups However, in contrast to the essential oil, few studies have in- received LAE at three different doses (857, 1714, and 3428 vestigated the medicinal properties of the aqueous extracts of mg/kg). The rats were placed in an acrylic cylinder (45 cm lavender. × 19.2 cm i.d.) filled with water at 25 ± 1℃ to a depth of The purpose of the present study was to examine the 17 cm for 15 min without any treatment (pre-test session). antidepressant-like effects of an aqueous extract of lavender Twenty-four hours later, the rats were exposed to the same (LAE) by using the forced swimming test (FST), the most conditions for 5 min (test session). Between the pre-test and widely used animal model of depression. test sessions, the test solutions were orally administered three times, the first just after the pre-test session, then 5 h before Material and Methods the test session, and finally 1 h before the test session. Time Animals 6-week-old male Wistar rats (110 − 130 g) spent by the rats in active movement and immobility was re- were obtained from Japan SLC (Shizuoka, Japan). The rats corded during the 5-min test session by a video camera. The were group-housed (seven rats/cage; cage size: 420 × 263 × tapes were evaluated by observers not informed of the treat- 199 mm height) in a room at a constant temperature (23 ± ment that each animal had received. After each test, the tank 1℃) and humidity (50 ± 5%), and a 12-hour light-dark cycle was refilled with fresh water. (light: 7:00 − 19:00). Controlled purified diets were prepared Open field test (OFT) procedure The locomotor activ- according to CE-2 (CLEA Japan, Inc., Shizuoka, Japan). The ity in a novel environment was recorded employing the OFT. animals consumed the diets and tap water ad libitum. All Fourteen male rats were divided into two groups (seven rats animals were handled daily for 7 days to familiarize them per group): one group served as the control and the other with the experimental equipment and to minimize nonspe- group received LAE at a dose of 3428 mg/kg. The test solu- cific stress responses during the experiment. Behavioral tests tions were orally administered three times, i.e., 24, 5, and 1 were performed between 13:00 and 17:00 h. A different set h before the test. The arena was a circular black box (70 cm of animals was used for each behavioral test. All experimen- diameter and 40 cm height). The test was begun by placing tal procedures were in accordance with the Guidelines of the the animals at the center of the arena. The test used a CCD University of Shizuoka for the Care and Use of the Labora- camera (2 frames/s) and a computer video tracking system tory Animals, based on those of the American Association Image OF (O'hara & Co., Ltd., Tokyo, Japan), modified soft- for Laboratory Animal Science. ware based on the public domain NIH Image program (i). Chemicals Imipramine hydrochloride was purchased Locomotion (cm) and percent center (%) were analyzed for from Nacalai Tesque (Kyoto, Japan). (-)-Linalool and rosma- 5 min. Defecation was estimated by counting the number of rinic acid were purchased from Sigma-Aldrich Japan (Tokyo, fecal boluses deposited in the arena. Japan). Apigenin was purchased from Wako Pure Chemi- High performance liquid chromatography (HPLC) An cal Industries (Osaka, Japan). tert-Butylhydroquinone was Agilent 1200 Series HPLC system equipped with a diode purchased from Kanto Chemicals (Tokyo, Japan). All other array detector (Agilent Technologies, Palo Alto, CA) and reagents were of the highest grade commercially available. a Capcell Pak C18 MG column (250 mm × 4.6 mm i.d.; Preparation of LAE One hundred grams of dried lav- particle size, 5 µm; Shiseido, Tokyo, Japan) were used. The ender (Lavandula angustifolia Mill.) flowers were extracted operating conditions were as follows: column oven tem- with 2 L of distilled water at 90℃ for 1 h. The extract sepa- perature, 40℃; mobile phase, a linear gradient from 100% rated from the residue was evaporated in vacuo and lyophi- solvent A (water, pH 2.5 adjusted with phosphoric acid/ace- lized to give 25 g of LAE. The LAE was stored in a desicca- tonitrile 90:10) to 100% solvent B (acetonitrile) in 30 min, tor at 4℃ until needed. held at 100% B for 5 min; flow rate, 1 mL/min; and injection Treatments LAE and imipramine hydrochloride were volume, 1 µL. The samples were filtered though a 0.45-µm dissolved in distilled water just before use. All solutions PTFE membrane filter (DISMIC-13HP, Toyo Roshi, Tokyo, were orally administered at a volume of 1.0 mL/100 g body Japan) prior to injection. Wavelengths of 210, 310, and 350 weight of the rat. The control group only received distilled nm were used for the quantification of linalool, rosmarinic water. acid, and apigenin aglycone, respectively, and the UV spectra FST procedure The FST was performed according to (wavelengths from 200 to 400 nm) plus retention time were the method of Porsolt et al., with some modifications (Por- used for their identification. Antidepressant-like Effects of Lavender 475 Determination of total apigenin glycosides in LAE To result of imipramine was not significant, the analysis was determine the total concentration of apigenin glycosides in stopped so that the overall FWER did not exceed a level of LAE, hydrolysis was performed by a method of Hertog et 0.05 (two-tailed).